• Title/Summary/Keyword: NSP4

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Nucleotide sequence analysis and expression of NSP4 gene of human rotaviruses isolated in Korea (국내에서 분리된 사람 로타바이러스의 NSP4 유전자 염기서열 분석 및 발현)

  • Jung, Dong-hyuk;Song, Yun-kyung;Kim, Kyung-mi;Park, Hyo-sun;Back, Myoung-soon;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.89-100
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    • 2002
  • The nonstructural glycoprotein NSP4, encoded by the 10th gene of rotavirus, has been known to play important roles in viral assembly and pathogenesis. The NSP4 genes of human rotavirus Korean isolates, designated as CBNU/HR-1, CBNU/HR-2, CBNU/HR-3, and CBNU/HR-4, were cloned, sequenced and characterized. Also, the NSP4 gene of the CBNU/HR-1 was expressed in a baculovirus-insect cell system. The sequence data indicated that the NSP4 genes of human rotavirus Korean isolates were 750 or 751 bases in length and encoded one open reading frame of 175 amino acids. Two glycosylation sites were recognized in the NSP4 gene of human rotavirus isolates tested. The NSP4 of CBNU/HR-1, CBNU/HR-3, and CBNU/HR-4 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype B viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype A viruses. However, the NSP4 of CBNU/HR-2 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype A viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype B viruses. The Sf9 cells infected with recombinant baculovirus, inserted with NSP4 gene of CBNU/HR-1, produced specific cytopathic effects and the expressed NSP4 was detected by immunofluorescence staining using NSP4-specific monoclonal antibody(MAb). The expressed NSP4 migrated at 16-26 kDa on SDS-PAGE and reacted with NSP4-specific MAb by Western blotting.

Nucleotide sequence analysis and expression of NSP4 gene of avian rotavirus (조류 로타바이러스의 NSP4 유전자 염기서열분석 및 발현)

  • Shin, In-ho;Lee, Seung-chul;Kim, Won-yong;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.207-214
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    • 2005
  • The nonstructural protein 4 (NSP4) of rotavirus encoded by gene 10, plays an important role in rotavirus pathogenicity. In this study, NSP4 gene of avian rotavirus (AvRV-1, AvRV-2) was analyzed and expressed using baculovirus expression system. The sequence data indicated that the NSP4 gene of AvRV-1 and AvRV-2 were 727 bases in length, encoded one open reading frame of 169 amino acids beginning at base 41 and terminating at base 550, and had two glycosylation sites. Nucleotide sequences of NSP4 gene of AvRV-1 and AvRV-2 exhibited a high degree of homology ($88.1{\pm}7.6%$) with avian rotaviruses, namely Ty1, Ty3 and PO-13. Phylogenetic analysis showed that AvRV-1 and AvRV-2 belonged to genotype NSP4[E], which is widely found in group A avian rotaviruses. The baculovirus-expressed NSP4 migrated at 20-28 kDa and reacted with NSP4-specific antiserum by FA and Western blot. Furthermore, it was found to be a glycoprotein by using tunicamycin, which is a specific inhibitor of N-linked glycosylation.

Cloning and Sequencing of Nonstructural Protein (NSP4) Gene of Bovine Rotavirus Isolated in Korea (국내분리 소 로타바이러스 NSP4 유전자의 크로닝 및 염기서열 분석)

  • Back, Myoung-Soon;Kim, Won-Yong;Kang, Shien-Young
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.1-10
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    • 2000
  • The nonstructural protein NSP4, encoded by gene 10 of rotavirus, has been shown to playa role in viral assembly and known to be an enterotoxin, causing diarrhea in mouse pups. NSP4 gene was cloned from CBNU-2 (virulent bovine rotavirus/diarrheic fecal sample) and CBNU-1 (cell-culture adapted bovine rotavirus/isolated from CBNU-2 and 75 times passaged on MA104 cells), respectively, by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequenced and compared. The sequence data indicated that the NSP4 genes of bovine rotavirus (BRV) were 751 bases in length and encoded one open reading frame of 175 amino acids beginning at base 42 and terminating at base 569. Differences in nucleotide sequence between CBNU-2 and CBNU-1 were observed at 6 positions (base 274, 296, 391, 394, 396 and 579). NSP4 gene of BRV exhibited a high degree of nucleotide (90% and 94%) and amino acid sequence (91% and 97%) homology with those of SA11 and UK but a low degree of nucleotide (77% and 79%) and amino acids sequence (81% and 85%) homology with those of Wa and OSU.

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Impact of Tofu Paste and Non-starch Polysaccharides on Oil Uptake Reduction in Cake Doughnuts (케이크 도넛의 흡유저감에 대한 두부 페이스트와 비전분성 탄수화물 고분자의 영향)

  • Jung, Gil-Young;Lee, Hyeon-Jeong;Ko, Eun-Sol;Kim, Hyun-Seok
    • Food Engineering Progress
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    • v.21 no.1
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    • pp.72-78
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    • 2017
  • The objective of this study was to investigate the effects of combinations of tofu paste and non-starch polysaccharides (NSP) on the oil uptake reduction (OTR) of deep-fat fried cake doughnuts. OTR agents were tofu paste (from grinding tofu with deionized water, followed by passage through a 60 mesh sieve), and five neutral and nine anionic NSPs. A control doughnut (without tofu paste or NSP), tofu doughnut (with tofu paste) and NSP-tofu doughnut (with tofu paste and NSP) were prepared. The moisture and total lipid (TL) content, cross-section image, color characteristic, and specific volume were measured. The tofu and NSP-tofu doughnuts exhibited higher moisture and lower TL content than the control. OTR was 10.8% for the tofu doughnut, and between 13.2% and 41.2% for the NSP-tofu doughnut. The highest OTR (41.2%) was found in the NSP-tofu doughnut with a combination of tofu paste and sodium alginate (NaA). The specific volume of the NSP-tofu doughnuts with combinations of tofu paste with NaA (2.5 mL/g), locust bean gum (2.5 mL/g), and ${\kappa}$-carrageenan (2.4 mL/g) was very close to that of the control (2.6 mL/g). Considering the OTR and specific volume of doughnuts, the combination of tofu paste and NaA would be most effective in reducing the oil uptake of doughnuts during deep-fat frying.

Effects of Nitrogen Application Levels on Grain Yield and Yield-related Traits of Rice Genetic Resources (질소비료 시비 수준이 벼의 수량 및 수량구성요소에 미치는 영향)

  • Tae-Heon Kim;Suk-Man Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.68 no.4
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    • pp.276-284
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    • 2023
  • Nitrogen is a major and essential macronutrient for plant growth and development. However, excessive nitrogen application can lead to ecological pollution or greenhouse gas emissions, consequently resulting in climate change. In this study, we used 153 genetic resources of rice to evaluate the effects of the levels of nitrogen application on grain yield and yield-related traits. Significant differences were noted in the yield and yield-related traits of genetic resources between two nitrogen application levels, namely, 4.5 kg/10a (NN: normal nitrogen condition) and 9.0 kg/10a (LN: low-nitrogen condition). Among the tested traits, days to heading (DTH), clum length (CL), grain yield per plant (GYP), number of panicles per plant (NPP), and number of spikelets per panicle (NSP) decreased by 1.8 to 17.9% when the nitrogen application levels decreased from NN to LN. The 1,000-grain weight (TWG) and percentage of ripened grain (PRG) increased by 2.6 to 11.2% under these conditions. Based on nitrogen application levels, two-way analysis of variance (ANOVA) demonstrated significant differences in GYP, NPP, and PRG but not in NSP and TGW. NPP exhibited negative correlations with NSP (-0.44) and TGW (-0.44), and TGW displayed a negative correlation with PRG (-0.34), whereas, GYP exhibited a positive correlation with PRG (0.37) and NSP (0.38). A similar pattern was recorded under the LN condition. NPP, TGW, and PRG were clustered as PA (principle axis) 1 under the LN condition by factor analysis. NSP and GYP were clustered as PA (principle axis) 2. These results demonstrated NPP and NSP as the primary factors contributing to the decrease in grain yield under LN conditions. In conclusion, we selected eight genetic resources that exhibited higher GYP under both NN and LN conditions with higher NPP or NSP. These genetic resources can be considered valuable breeding materials for the adaptation of plants to nitrogen deficiency.

Deuterium-labeling Toward Robust Function of Organic Molecules: Enhanced Photo-stability of Partially Deuterated 1', 3', 3'-Trimethyl-6-nitrospiro[2H-1- benzopyran-2, 2'-indoline]

  • Kawanishi, Yuji;Inoue, Kyoko;Ohta, Shin-Ichi;Miyazawa, Akira
    • Rapid Communication in Photoscience
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    • v.3 no.4
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    • pp.64-66
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    • 2014
  • Synthesis of a deuterium-labeled derivative of nitrospirobenzopyran (NSP), one of representative photochromic compounds, has been described. Four deuteriums were successfully introduced on 1-methyl and ${\alpha}$-methyne relative to spiro-carbon in the title compound with more than 95atom%D purity. Main photodegraded products of NSP were two oxindoles in acetonitrile, and additional products were formed in poly(isobutyl-methacrylate) films possibly due to restricted molecular motion in polymer matrix. Quantitative HPLC analysis revealed that partial introduction of deuterium to NSP brought a noticeable isotope effect, recognizable enhancement in photo-resistivity of NSP, i.e.,8.3% in solutions and 29% in polymeric films.

Anti-inflammatory Activities of Ethanol Extracts from Leaf, Seed, and Seedpod of Nelumbo nucifera (연잎, 연자육, 연자방 에탄올 추출물의 항염증 활성)

  • Lee, Eun-Joo;Seo, Yu-Mi;Kim, Yong-Hyun;Chung, Chungwook;Sung, Hwa-Jung;Sohn, Ho-Yong;Park, Jong-Yi;Kim, Jong-Sik
    • Journal of Life Science
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    • v.29 no.4
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    • pp.436-441
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    • 2019
  • Nelumbo nucifera, also known as sacred lotus, has mainly been used as a food throughout the Asian countries. In the present study, we prepared ethanol extracts from leaf (NL), seed (NS), and seedpod (NSP) of Nelumbo nucifera and investigated their anti-inflammatory activities in mouse macrophage RAW 264.7 cells. To evaluate the anti-inflammatory activities of NL, NS, and NSP, nitric oxide (NO) production was measured in LPS-stimulated RAW 264.7 cells. NL, NS, and NSP significantly reduced NO production in a dose-dependent manner without affecting cell viabilities. NL, NS, and NSP dramatically decreased the protein expression of pro-inflammatory genes such as iNOS and COX-2. NL, NS, and NSP also suppressed phosphorylation of MAPKs and the nuclear translocation of $NF-{\kappa}B$ p65 indicating they have their anti-inflammatory activities via regulating mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B ($NF-{\kappa}B$) pathways. In addition, we analyzed the production of reactive oxygen species (ROS) by the treatment of NL, NS, and NSP. All extracts reduced ROS production in a dose-dependent manner. And also, they increased heme oxygenase-1 (HO-1) protein expression and the nuclear translocation of nuclear respiratory factor 2 (Nrf2). In conclusion, our results suggest that Nelumbo nucifera has its anti-inflammatory activity via regulating MAPKs, $NF-{\kappa}B$, and Nrf2/HO-1 pathways.

Molecular characterization of avian rotavirus isolated in Korea

  • Wang, Jun-Hui;Koo, Bon-Sang;Mo, In-Pil;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.36 no.1
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    • pp.23-30
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    • 2013
  • An avian rotavirus (AvRV-2) was isolated from feces of broilers suffering from acute gastroenteritis in 2011. It was the first avian rotavirus isolated in Korea. To investigate the molecular characteristics of AvRV-2, the VP4, VP6, VP7 and NSP4 gene nucleotide sequences were determined and compared with those of rotavirus strains available in the GenBank database. The phylogenetic tree of VP7 gene showed that AvRV-2 had a high degree of nucleotide sequence homology (93.4% to 94.7%) with those of rotaviruses belonging to genotype G19 cluster. The phylogenetic tree of the VP4 gene revealed a high degree of nucleotide sequence homology (95.8% to 95.9%) with genotype P[30] rotaviruses isolated from chickens. The VP6 and NSP4 gene nucleotide sequences showed the highest identities with those of avian strains with 95.3% to 96.4% and 90.3% to 92.2%, respectively. Genetic characterization of the VP4, VP6, VP7 and NSP4 showed that AvRV-2 strain was most closely related to chicken rotavirus strains from Germany and Japan. Comparative nucleotide sequences and phylogenetic analysis indicated that avian rotavirus isolated from broilers belonged to genotype G19P[30] and it was the first report on avian rotavirus infection in Korea.

Comparison of the Properties of Wheat Flours Supplemented with Various Dietary Fibers

  • Lee, Hyun-Ju;Shin, Mal-Shick
    • Food Science and Biotechnology
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    • v.15 no.5
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    • pp.746-751
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    • 2006
  • The effects of resistant starch (RS) and non-starch polysaccharide (NSP) addition on the properties of hard wheat flour were investigated. Total dietary fiber (TDF) levels of various NSP ranged from 78.3-100.0%, but TDF and RS levels of autoclaved RS3 and cross-linked RS4 were 16.1 and 35.0% and 13.2 and 90.9%, respectively. DF-supplemented flour increased swelling power, but RS4-supplemented flour exhibited the lowest it. Solubility increased with the addition of pectin and RS3, but decreased with the addition of cellulose and RS4. RS-supplemented flour had increased lightness (L), but decreased values of redness (a) and yellowness (b). RS3 and pectin increased the dough development time, but RS4, cellulose, and chitosan decreased it. The water absorptions of pectin- and RS4-supplemented flours increased, however the dough stability decreased. The initial pasting temperatures of RS- and NSP-supplemented flours increased regardless of amount added, but the maximum peak viscosity decreased for all except the pectin-supplemented flour.

Genetic characterization of bovine rotavirus isolates in Korea

  • Yang, Dong-Kun;Kim, Byoung-han;Lee, Kyung-Woo;Kim, Yeon-Hee;Song, Jae-Young;Park, Joong-Won;Son, Seong-Wan
    • Korean Journal of Veterinary Research
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    • v.48 no.4
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    • pp.423-429
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    • 2008
  • Throughout the world, rotavirus infections cause extensive morbidity in human infants and diarrhea in animals such as white scour caused by bovine rotavirus in calves. We isolated three rotavirus strains designated KV0407, KV0418, and KV0426 from 103 fecal samples of diarrheic calves. The genes coding for proteins VP4, VP6, VP7, and NSP4 from strain KV0407 were sequenced and compared with the nucleotide sequences of other known strains of rotavirus. The KV0407 VP4 gene was highly homologous to the OSU (99.4%) and JL94 (99.4%), but not the B223 (62.4%) and K33 (62.4%) VP4 genes. The KV0407 and KV0418 VP7 genes were most similar to the OSU and super-short type VMRI VP7 genes. Based on nucleotide sequence analysis, the KV0407 strain was tentatively assigned to A serogroup (SG I), G5P[7], NSP4 genotype B and the KV0418 and KV0426 strains were assigned to A serogroup (SG I), G6P[5], NSP4 genotype A. The genetic characterization of these bovine rotavirus isolates could be useful for the diagnosis and prevention of diarrhea in calves.